Several publications and patent documents are cited throughout the specification in order to describe the state of the art to which this invention pertains. Each of these citations is incorporated herein by reference as though set forth in full.
Non-viral gene delivery is a critically important method of delivering genes to a cell both in vitro and in vivo. To enhance the transfer of DNA into the cells, polycations and polymer agents have been employed that can 1) bind and condense DNA, 2) protect the DNA from degradation, and 3) enhance transport of the DNA into the cell (Wagner et al. (1990) PNAS, 87:3410-3414; Kabanov et al. (1993) Bioconj. Chem., 4:448-454; Boussif et al. (1995) PNAS, 92:7297-7301; Tang et al. (1997) Gene Ther., 4:823-832; Pollard et al. (1998) J. Biol. Chem., 273:7507-7511; Godbey et al. (1999) PNAS, 96:5177-5181; Merdan et al. (2002) Pharm. Res., 19:140-146). Amphiphilic block copolymers have been employed to increase the transfer of naked DNA in vivo into a variety of tissues including muscle and skin tissues and tumors (Lemieux et al. (2000) Gene Ther., 7:986-991; Liaw et al. (2001) Gene Ther., 8:999-1004; Alakhov et al. (2001) Expert Opin. Biol. Ther., 1:583-602; Gebhart et al. (2003) Controlled Release Society, Glasgow, Scotland, UK; Pitard et al. (2002) Human Gene Ther., 13:1767-1775; Batrakova et al. (2003) J. Pharmacol. Exp. Ther., 304:845-854). Additionally, amphiphilic block copolymers have been shown to increase the transfer of polycation-DNA complexes (Nguyen et al. (2000) Gene Ther., 7:126-138; Gebhart et al. (2002) Bioconj. Chem., 13:937-944; Astafieva et al. (1996) FEBS Lett., 389:278-280; Kuo, J. H. (2003) Biotechnol. Appl. Biochem., 37:267-271).
Transduction of cells with viral vectors, such as adenoviral and lentiviral, is also increased in the presence of amphiphilic block copolymers (March et al. (1995) Hum. Gene Ther., 6:41-53; Feldman et al. (1997) Gene Ther., 4:189-198; Van Belle et al. (1998) Hum. Gene Ther., 9:1013-1024; Maillard et al. (2000) Gene Ther., 7:1353-1361; Dishart et al. (2003) J. Mol. Cell. Cardiol., 35:739-748).
All of the above studies demonstrate the ability of amphiphilic block copolymers to assist in the transfer of DNA and DNA-containing compositions into cells. However, such studies are silent as to the effect of the polymers on gene expression of genes already present in the cell.